Barcoded nanoparticles for high throughput in vivo discovery of targeted therapeutics
Barcoded nanoparticles for high throughput in vivo discovery of targeted therapeutics
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DOI：
10.1073/pnas.1620874114
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发表时间：
2017-02-21
影响因子：
11.1
通讯作者：
Wang, Eric T.
中科院分区：
文献类型：
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作者：
Dahlman, James E.;Kauffman, Kevin J.;Wang, Eric T.
核酸治疗剂受限于向靶组织和细胞的低效递送以及对纳米颗粒结构如何影响向脱靶器官的生物分布的不完全理解。尽管已经设计了数千种纳米颗粒制剂来递送核酸，但是大多数纳米颗粒已经在细胞培养环境中进行了测试，其不再现全身性体内递送。为了增加可以在体内测试的纳米颗粒的数量，我们开发了一种方法来同时测量许多化学上不同的纳米颗粒的生物分布。我们配制了携带特定核酸条形码的纳米颗粒，管理了颗粒池，并通过对条形码进行深度测序来量化颗粒的生物分布。该方法区分了先前表征的肺靶向和肝靶向纳米颗粒，并准确报告了递送至组织的核酸的相对量。条形码序列不影响递送，并且对于供试颗粒，未观察到颗粒混合的证据。通过同时测量30种纳米颗粒在8种组织中的生物分布，我们确定了促进某些组织相对于其他组织递送的化学性质。最后，当与siRNA一起配制时，分布到肝脏的颗粒也沉默了肝细胞中的基因表达。该系统可以促进靶向特定组织和细胞的纳米颗粒的发现，并加速化学结构与体内递送之间关系的研究。
Nucleic acid therapeutics are limited by inefficient delivery to target tissues and cells and by an incomplete understanding of how nanoparticle structure affects biodistribution to off-target organs. Although thousands of nanoparticle formulations have been designed to deliver nucleic acids, most nanoparticles have been tested in cell culture contexts that do not recapitulate systemic in vivo delivery. To increase the number of nanoparticles that could be tested in vivo, we developed a method to simultaneously measure the biodistribution of many chemically distinct nanoparticles. We formulated nanoparticles to carry specific nucleic acid barcodes, administered the pool of particles, and quantified particle biodistribution by deep sequencing the barcodes. This method distinguished previously characterized lung-and liver-targeting nanoparticles and accurately reported relative quantities of nucleic acid delivered to tissues. Barcode sequences did not affect delivery, and no evidence of particle mixing was observed for tested particles. By measuring the biodistribution of 30 nanoparticles to eight tissues simultaneously, we identified chemical properties promoting delivery to some tissues relative to others. Finally, particles that distributed to the liver also silenced gene expression in hepatocytes when formulated with siRNA. This system can facilitate discovery of nanoparticles targeting specific tissues and cells and accelerate the study of relationships between chemical structure and delivery in vivo.